KR20060002778A - Improved antitumoral treatments - Google Patents

Abstract

Aplidine and aplidine analogues are of use for the treatment of cancer, in particular in the treatment of leukemias and lymphomas, especially in combination therapies.

Description

Improved antitumoral treatments

The present invention relates to the combination of aplidines or aplidine analogs with other antitumor agents, the use of these combinations in the treatment of cancer, in particular in the treatment of leukemias and lymphomas.

Oh, Dean Shipley (aplidine) (Lodi demnin dehydrogenase B: Dehydrodidemnin B) is a Mediterranean marine current Pinang (tunicate) O Plymouth Inn Stadium albicans (Aplidium albicans ) and is a cyclic depsipeptide, the subject of WO 9109485. It is associated with a compound known as didemnin and has the following structure:

More information regarding aplidines, aplidine analogs, their use, formulation and synthesis can be found in patent applications WO 98 1352, WO 99 42125, WO 01 76616, WO 01 35974, WO 02 30441 and WO 02 02596. have. We incorporate the contents of each of these PCR texts by specific reference herein.

In animal and human pre-ginseng studies and in Phase I clinical studies, the drug has been shown to have cytotoxic capacity against a wide range of tumor types, including leukemia and lymphoma. For example, Faircloth, G. et al.:"Dehydrodidemnin B (DDB) a new marine derived anticancer agent with activity against experimental tumour models ", 9th NCI-EORTC Symp New Drugs Cancer Ther (March 12-15, Amsterdam) 1996 , Abst 111; Faircloth, G. et al .: "Preclinical characterization of aplidine, a new marine anticancer depsipeptide", Proc Amer Assoc Cancer Res 1997, 38: Abst 692; Depenbrock H, Peter R, Faircloth GT, Manzanares I, Jimeno J, Hanauske AR.:"In vitro activity of aplidine, a new marine-derived anti-cancer compound, on freshly explanted clonogenic human tumour cells and haematopoietic precursor cells "Br. J. Cancer, 1998; 78: 739-744; Faircloth G, Grant W, Nam S, Jimeno J, Manzanares I, Rinehart K .: "Schedule-dependency of aplidine, a marine depsipeptide with antitumor activity" ', Proc. Am. Assoc. Cancer Res. 1999; 40: 394; Broggini M, Marchini S, D'Incalci M, Taraboletti G, Giavazzi R, Faircloth G, Jimeno J .: "Aplidine blocks VEGF secretion and VEGF / VEGF-R1 autocrine loop in a human leukemic cell line", Clin Cancer Res 2000; 6 (suppl): 4509; Erba E, Bassano L, Di Liberti G, Muradore I, Chiorino G, Ubezio P, Vignati S, Codegoni A, Desiderio MA, Faircloth G, Jimeno J and D'Incalci M .: "Cell cycle phase perturbations and apoptosis in tumour cells induced by aplidine ", Br J Cancer 2002; 86: 1510-1517; Paz-Ares L, Anthony A, Pronk La Twelves C, Alonso S, Cortes-Funes H, Celli N, Gomez C, Lopez-Lazaro L, Guzman C, Jimeno J, Kaye S .: "Phase I clinical and pharmacokinetic study of aplidine, a new marine didemnin, administered as 24-hour infusion weekly "Clin. Cancer Res. 2000; 6 (suppl): 4509; Raymond E, Ady-Vago N, Baudin E, Ribrag V, Faivre S, Lecot F, Wright T, Lopez Lazaro L, Guzman C, Jimeno J, Ducreux M, Le Chevalier T, Armand JP .: "A phase I and pharmacokinetic study of aplidine given as a 24-hour continuous infusion every other week in patients with solid tumor and lymphoma ", Clin. Cancer Res. 2000; 6 (suppl): 4510; Maroun J, Belanger K, Seymour L, Soulieres D, Charpentier D, Goel R, Stewart D, Tomiak E, Jimeno J, Matthews S .: "Phase I study of aplidine in a 5 day bolus q 3 weeks in patients with solid tumors and lymphomas ", Clin. Cancer. Res. 2000; 6 (suppi): 4509; Izquierdo MA, Bowman A, Martinez M, Cicchella B, Jimeno J, Guzman C, Germa J, Smyth J .: "Phase I trial of aplidine given as a 1 hour intravenous weekly infusion in patients with advanced solid tumors and lymphoma", Clin . cancer Res. 2000; 6 (suppl): 4509.

Mechanistic studies have shown that aplidine can block VEGF secretion in ALL-MOLT4 cells, and is at low concentration (5 nM) in AML and ALL samples from pediatric patients with de novo or recurrent ALL and AML. In vitro cytotoxicity was observed. Aplidine appears to induce G1 and G2 arrest in drug treated leukemia cells in vitro. Except for down-regulation of the VEGF receptor, little is known about the mode of action of aplidine.

In a Phase 1 clinical study with aplidine, L-carnitine was given or co-administered as a 24-hour pretreatment to prevent myelotoxicity. See WO 02 30441. It has been found that co-administration of L-carnitine may improve the recovery of the drug induced muscle toxicity, thereby enabling a dose increase of aplidine.

Thus, in a Phase 1 study, aplidine was not bone marrow toxic at the maximum tolerated dose, except for weak lymphopenia. These properties make aplidine a potentially useful drug for the treatment of leukemia. Adding aplidine to current chemotherapy for leukemia can improve efficacy without reducing the dose of drug recognized to have anti-leukemic activity due to increased myelotoxicity. This is particularly relevant for the treatment of relapsed ALL and newly diagnosed and relapsed AML, a disease with a relatively poor prognosis, currently being treated with a myeloid toxic drug combination.

The inventors have established for the first time that aplidine and aplidine analogues can enhance other anticancer agents and thus be successfully used in combination therapy for the treatment of cancer. The present invention relates to pharmaceutical compositions, pharmaceutical dosage forms, kits and methods for the treatment of cancer using these combination therapies.

According to one embodiment of the present invention, the present inventors provide an effective combination therapy based on aplidine and aplidine analogs using other drugs effective for the treatment of cancer. Preferably, said other drug is effective in the treatment of leukemia and / or lymphoma. Most preferably, the other drug is selected from the group consisting of methotrexate, cytosine arabinoside, mitoxantrone, vinblastine, methylprednisolone and doxorubicin.

In another embodiment, the present invention is a method of treating primary and / or metastatic cancer, wherein the therapeutically effective amount of aplidine or aplidine analog, or a pharmaceutically acceptable prodrug thereof, in a patient in need of such treatment , Salts, solvates or hydrates, and other drugs or pharmaceutical agents in a therapeutically effective amount effective for the treatment of the cancer administered prior to, during or after administration of the aplidine or aplidine analogues. And a method comprising administering an acceptable prodrug, salt, solvate or hydrate thereof.

Preferably, said other drug is effective in the treatment of leukemia and / or lymphoma. Most preferably, the other drug is selected from the group consisting of methotrexate, cytosine arabinoside, mitoxantrone, vinblastine, methylprednisolone and doxorubicin. The other drug may form part of the same composition or may be prepared as a separate composition for administration at the same time or at different times.

The cancer to be treated is preferably leukemia or lymphoma, most preferably ALL, AML, CML, MML or CLL.

In another aspect, the invention comprises administering to a patient in need thereof an amount of an aplidine or an aplidine analog, or a pharmaceutically acceptable prodrug, salt, solvate, or hydrate thereof, wherein the treatment of cancer Drugs effective in the treatment of leukemia and / or lymphoma, preferably drugs selected from the group consisting of methotrexate, cytosine arabinoside, mitoxantrone, vinblastine, methylprednisolone and doxorubicin, Or a method of increasing the efficacy of a pharmaceutically acceptable prodrug, salt, solvate or hydrate thereof. Aplidine or the aplidine analog is administered before, during or after administration of the other drug.

Aplidines or aplidine analogs can increase the therapeutic efficacy of some cancer drugs. In one aspect, the result is synergistic rather than additive. Such synergistic combinations represent a preferred aspect of the present invention. Synergism can be represented using the Chou-Talalay method, or other methods. In another example, antagonism may be found.

In another aspect, the present invention includes a pharmaceutical composition comprising an aplidine or an aplidine analog, or a pharmaceutically acceptable prodrug, salt, solvate or hydrate thereof, and another drug effective for the treatment of cancer. do. Preferably, said other drug is effective in the treatment of leukemia and / or lymphoma. Most preferably, it is selected from the group consisting of said other drug methotrexate, cytosine arabinoside, mitoxantrone, vinblastine, methylprednisolone and doxorubicin.

The invention also relates to aplidines or aplidine analogs, or pharmaceutically acceptable prodrugs, salts, solvates or hydrates, dosage forms, other drugs effective for the treatment of cancer, or pharmaceutically acceptable prodrugs thereof. , Kits for use in the treatment or prevention of cancer, including instructions for the use of each agent in a dosage form of a salt, solvate or hydrate, and in combination for the treatment or prevention of cancer. Preferably, said other drug is effective in the treatment of leukemia and / or lymphoma. Most preferably, it is selected from the group consisting of said other drug methotrexate, cytosine arabinoside, mitoxantrone, vinblastine, methylprednisolone and doxorubicin.

In another aspect, the present invention relates to the use of aplidine for the treatment of chronic lymphocytic leukaemia.

Cancer is meant to include tumors, neoplasia and any other malignant tissue or cell. The present invention generally relates to the use of aplidines or aplidine analogs in combination for the treatment of cancer, but most preferably for the treatment of other leukemias and lymphomas.

In order to study the possible fortification of other anticancer agents with aplidine, we have initiated a systematic study of drug combinations for possible use in leukemias and lymphomas. Aplidine has been found to be an effective in vitro cytotoxic agent against fresh cells obtained from six chronic lymphocytic leukemia (CLL) patients as well as against primary cells obtained from patients with preB-ALL (DM4). The IC ₅₀ value was 10 nM for 3 days exposure to DM4 predominantly and after 11 days exposure with primary CLL samples.

Drug combination studies were performed on established cell lines rather than primary cells. We studied three cell lines, namely K562, CCRF-CEM and SKI-DLCL, representing acute leukemia, lymphocytic lymphoma and diffuse B cell large cell lymphoma. According to the data of the examples, aplidine lowers the drug's IC _50, thereby reducing the drug's IC ₅₀ and the effects of methotrexate, cytosine arabinoside, mitoxantrone, vinblastine, methylprednisolone, as well as doxorubicin in K562, CCRF-CEM and SKI-DLCL cells. To strengthen.

Therefore, the inventors have found that aplidine is a potent cytotoxic agent against cells of various hematological cancers. Importantly, we first established that aplidine inhibits the growth of CLL cells in culture. The inventors also found that aplidine is effective in leukemia, such as methotrexate (MTX), cytosine arabinoside (AraC), mitoxantrone (Mitox), vinblastine (Vinb), methylprednisolone (Metpred) and doxorubicin (DOX) It has been found to enhance the cytotoxicity of agents used for treatment.

Leukemia is classified according to how fast it progresses. Acute leukemia can grow quickly and overwhelm the body in weeks or months. In contrast, chronic leukemia grows slowly and progressively worsens over the years.

Hematopoietic (hematopoietic) cells of acute leukemia remain immature, proliferating and accumulating very quickly. Therefore, acute leukemia needs to be treated immediately or it can be fatal within a few months. Fortunately, some subtypes of acute leukemia can respond to and cure available treatments. Children often develop acute forms of leukemia and are treated differently from leukemia in adults.

In chronic leukemia, the blood producing cells eventually grow or differentiate, but are not "normal". These cells remain in the bloodstream longer than normal white blood cells and do not fight infection well.

Leukemias are also classified according to the type of leukocytes that proliferate: lymphocytes (immune system cells), granulocytes (bacterial disrupting cells) or monocytes (macrophage forming cells). When the abnormal cell leukocytes are primary granulocytes or monocytes, the leukemia is classified as myeloid or myeloid leukemia. On the other hand, when the abnormal cells are derived from myeloid lymphocytes, the cancer is classified as lymphocyte leukemia.

Other cancers known as lymphomas arise from lymphocytes in the lymph nodes, pancreas and other organs. Such cancers do not originate in the bone marrow and have different biological behaviors than lymphocytic leukemia.

There are dozens of other leukemias, but the four types occur most frequently. These classifications were based on whether the leukemia was acute versus chronic and myelogenous versus lymphocytic. In other words:

Acute myeloid leukemia (AML) (acute myelogenous leukemia) : In addition, it is also known as acute non-lymphocytic leukemia (ANLL), the most common form of adult leukemia. Most patients are of retirement age (mean age at diagnosis = 65 years) and more men develop than women. Fortunately, due to recent developments in therapies, AML can remain relieved (reduction of disease) in about 60% to 70% of adults receiving adequate treatment. Initial response rates are about 65-75% but overall treatment rates are higher with orders of 40-50%.

Chronic myeloid leukemia ( CML ) is known as myelodysplasia. That is, a disease in which bone marrow cells proliferate (double) outside bone marrow tissue. CML is easy to diagnose because it has a genetic specificity, or marker, that can be readily identified under a microscope. About 95% of CML patients have a genetic translocation between chromosomes 9 and 22 in their leukemia cells. Philadelphia chromosomes cause uncontrolled reproduction and proliferation of all forms of white blood cells and platelets (blood coagulation factors). CML is not yet treatable by standard chemotherapy or immunotherapy.

Acute lymphocytic leukemia (ALL) (Acute lymphocytic leukemia) : In addition, the acute lymphocytic leukemia, and hair bulb is known as (acute lymphoblastic leukemia), initial bigwa miliary white blood cells, or the malignant disease caused by the over growth and development of lymphocytes. The leukemia originates in the bone marrow (B-cells), thymus (T-cells) and blast cells of lymph nodes. ALL occurs most often at 4 years of age, mainly in children.

Chronic lymphocytic leukemia (CLL) (Chronic lymphocytic leukemia) is the most common leukemia in North America and Europe. CLL is a disease of older adults and very rare among people under 50. Men with CLL average 2: 1 more than women. CLL is believed to result from the gradual accumulation of mature, long-lived lymphocytes.

Therefore, this cancer is not caused by overgrowth as much as the extreme lifespan and build-up of malignant cells. Although the rate of accumulation varies from subject to subject, widespread tumor burden eventually leads to complications in all CLL patients.

The compositions of the present invention may comprise both ingredients (drugs) in a single pharmaceutically acceptable formulation. Alternatively, the components may be formulated separately and administered in combination with each other. Various pharmaceutically acceptable formulations well known to those skilled in the art can be used in the present invention. Selection of a suitable formulation for use in the present invention can be routinely performed by one skilled in the art based on the mode of administration and solubility characteristics of the components of the composition.

Examples of pharmaceutical compositions comprising aplidine or aplidine analogs include liquids (solutions, suspensions or emulsions) having a composition suitable for intravenous administration, wherein the composition is selected from the neat compound or any carrier or other pharmacologically It can be included in combination with an active compound. Dissolved aplidine is substantially degraded under heat and light stress test conditions, and lyophilized dosage forms have been developed. See WO 99/42125, incorporated herein by reference.

Administration of aplidine or a composition of the present invention is based on the Dosing Protocol, preferably by intravenous infusion. We prefer that an infusion time of up to 72 hours be used, more preferably 1 to 24 hours, most preferably about 1 hour, about 3 hours or about 24 hours. Particular preference is given to short infusion times which allow the treatment to be carried out without overnight stay in the hospital. However, if necessary, the infusion can be about 24 hours or more. Infusion can vary in various aspects, eg once a week, twice a week, three times a week, or more frequently a week, optionally repeated weekly with a weekly gap. Aspects can be performed at suitable intervals.

The correct dosage of the compounds of the combination will depend on the particular formulation, method of application, and particular location, host and tumor to be treated. Other factors can be considered such as age, weight, sex, food, time of administration, rate of secretion, conditions of the host, drug combination, sensitivity of the reaction and the severity of the disease. Administration can be carried out continuously or periodically within the maximum tolerated dose. Further guidance on the administration of aplidine is given in WO 0135974, which is incorporated herein by reference in its entirety.

For the present invention, analogs of aplidine can be used in place of APL, aplidine itself. Such compounds are generally as defined in WO 0202596. Examples of compounds for the present invention include preferred compounds given in WO 0202596, and we introduce discussions of preferred compounds given in WO0202596 and related aspects in the context of the present invention. More preferably, the analogs are structurally similar to aplidine and are usually different from aplidine in terms of one amino acid or terminal side chain. The other amino acid may be in the ring portion or side chain of the molecule. Many examples of such compounds are given in WO0202596, and they are candidates for use in the present invention.

Aplidine inhibits growth of CLL cells in culture.

Aplidine is a potent inhibitor of preB-ALL cells in culture.

Figure 3. Cytotoxic dose-response curves of CCRF-CEM (A), SKI-DLCL (B) and K562 (C) after 96 hours of aplidine treatment.

Figure 4 shows the Chou-Talalay analysis of the combination of aplidine and AraC in CCRF-CEM cells.

5 shows Chou-Talalay analysis of the combination of aplidine and AraC in SKI-DLCL cells.

Figure 6 shows the Chou-Talalay analysis of the combination of aplidine and mitoxantrone in CCRF-CEM cells.

7 shows Chou-Talalay analysis of the combination of aplidine and mitoxantrone in SKI-DLCL cells.

Figure 8. Chou-Talalay analysis of the combination of aplidine and methotrexate in CCRF-CEM cells.

Figure 9 shows the Chou-Talalay analysis of the combination of aplidine and doxorubicin in CCRF-CEM cells.

10. Chou-Talalay analysis of the combination of aplidine and vinblastine in CCRF-CEM cells.

Figure 11 shows the Chou-Talalay analysis of the combination of aplidine and doxorubicin in SKI-DLCL cells.

12. Chou-Talalay analysis of the combination of aplidine and vinblastine in SKI-DLCL cells.

Figure 13 shows the Chou-Talalay analysis of the combination of aplidine and methylprednisolone in SKI-DLCL cells.

14. The combination of IC ₂₀ of aplidine lowered IC ₅₀ of AraC in CCRF-CEM (FIG. 12A) and SKI-DLCL (FIG. 12B) cells after 96 hours of incubation.

15. The effect of aplidine on tumor size in vivo in combination with a single agent and AraC.

Example  One

Aplidine was tested on various primary cells obtained from patients with hematological cancer. The cells used were as follows:

Fresh cells from 6 chronic lymphocytic leukemia patients

Primary cells obtained from patients with preB-ALL (DM4)

Patient samples were obtained with informed consent, and CLL cells were isolated by concentration gradient centrifugation on a histopaque. The medium used was RPMI supplemented with 10% autologous serum and L-glutamine. Cultures were incubated with 10 nM aplidine and cell viability was measured at days 3, 7, 11 and 18 and compared to the viability of untreated cells and STI 571 (0.5 mM).

The results of these studies are shown in FIGS. 1-2.

Example  2

In order to study possible fortification of other anticancer agents, we conducted a study of drug combinations usable in leukemias and lymphomas.

Drug combination studies were performed on established cell lines rather than primary cells. We studied three cell lines, namely K562, CEM for acute lymphocytic leukemia and SKI-DLCL for diffuse large cell lymphoma as a model for acute myeloid leukemia.

IC ₂₀ of aplidine with dose ranges of methotrexate, cytosine arabinoside and doxorubicin And a combination study consisting of IC ₅₀ was tested to determine if aplidine can enhance the effects of these drugs.

The results are shown in Table 1:

p <0.01, p <0.05, p <0.05

Clearly, these data show that aplidine enhances the effects of doxorubicin, methotrexate and cytosine arabinoside by significantly reducing the IC ₅₀ for the drug.

Example 3 In Vitro Study to Determine the Effect of Aplidine as a Single Agent on CCRF-CEM, SKI-DLCL, and K562 Cell Lines

CCRF-CEMS, SKI-DLCL and K562 cells were maintained in RPMI 1640 supplemented with 10% FCS. To determine the cytotoxicity of aplidine against all cell lines and to obtain the IC ₅₀ of aplidine in these cell lines, the cells were plated in 96 well plates and 96 hours in an incubator that was moistened with 5% CO ₂ . Incubated for Cell viability was measured by XTT assay in an automated plate reader. We found that aplidine is cytotoxic to all cell lines at an IC ₅₀ dose of 0.5-1.0 nM (FIG. 3).

Example 4 In Vitro Effect of Aplidine + Drug Combination at Fixed Dose of IC ₅₀ : IC ₅₀ for All Cell Lines

Methotrexate, cytosine arabinoside C (ara-C), mitoxantrone, methylprednisolone, vinblastine and doxorubicin were tested in combination with aplidine.

Chou-Talalay assay was used to analyze the drug combination. If the Combination Index (CI) obtained by this assay is less than 1, the drug is synergistic; If CI is 1, the drug is additive; If the CI is greater than 1, the drug is antagonistic.

All cytotoxicity studies were performed by using XTT or MTS. We first determined IC ₅₀ doses of these drugs in SKI-DLCL, CCRF-CEM and K562 cell lines. We investigated drug combinations using a fixed ratio of IC50 (Aplidine): IC50 (Drug X).

Table 2 shows the combination of aplidine and Ara-C at a (IC50: IC50) dose in CCRF-CEM cells.

Table 2.

Chou-Talalay analysis of the aplidine and AraC combination in CCRF-CEM cells can be seen in FIG. The CI for this combination in CCRF-CEM cells is 0.469.

Table 3 shows the combination of aplidine and Ara-C at the (IC50: IC50) dose in SKI-DLCL cells.

Table 3.

The Chou-Talalay assay results of the combination of aplidine and AraC in SKI-DLCL cells can be seen in FIG. 5. The CI for this combination in SKI-DLCL cells is 0.306.

Table 4 shows the combination of aplidine and Ara-C at the (IC50: IC50) dose in K562 cells.

Table 4.

The CI for this combination in K562 cells is 0.502.

Table 5 shows the combination of aplidine and mitoxantrone at (IC50: IC50) doses in CCRF-CEM cells.

Table 5.

The Chou-Talalay analysis of the aplidine and mitoxantrone combinations in CCRF-CEM cells can be seen in FIG. 6. The CI for this combination in CCRF-CEM cells is 0.911.

Table 6 shows the combination of aplidine and mitoxantrone at (IC50: IC50) doses in SKI-DLCL cells.

Table 6.

Chou-Talalay analysis of the aplidine and mitoxantrone combinations in SKI-DLCL cells can be seen in FIG. 7. The CI for this combination in SKI-DLCL cells is 0.646.

Table 7 shows the combination of aplidine and mitoxantrone at doses (IC50: IC50) in K562 cells.

Table 7.

The CI for this combination in K562 cells is 0.487.

Table 8 shows the combination of aplidine and methotrexate at a (IC50: IC50) dose in CCRF-CEM cells.

Table 8.

The Chou-Talalay analysis of the aplidine and methotrexate combinations in CCRF-CEM cells can be seen in FIG. 8. The CI for this combination in CCRF-CEM cells is 0.950.

The results of Chou-Talalay analysis of the aplidine and doxorubicin combinations in CCRF-CEM cells can be seen in FIG. 9. The CI for this combination in CCRF-CEM cells is 1.952.

The Chou-Talalay assay results of the combination of aplidine and vinblastine in CCRF-CEM cells can be seen in FIG. 10. The CI for this combination in CCRF-CEM cells is 2.046.

Table 9 shows the combination of aplidine and doxorubicin at the (IC50: IC50) dose in SKI-DLCL cells.

Table 9.

The Chou-Talalay assay results of the combination of aplidine and doxorubicin in SKI-DLCL cells can be seen in FIG. 11. The CI for this combination in SKI-DLCL cells is 0.478.

Table 10 shows the combination of aplidine and vinblastine at the (IC50: IC50) dose in SKI-DLCL cells.

Table 10

The Chou-Talalay assay results of the combination of aplidine and vinblastine in SKI-DLCL cells can be seen in FIG. 12. The CI for this combination in SKI-DLCL cells is 0.760.

Table 11 shows the combination of aplidine and methylprednisolone at the (IC50: IC50) dose in SKI-DLCL cells.

Table 11.

The Chou-Talalay analysis of the aplidine and methylprednisolone combinations in SKI-DLCL cells can be seen in FIG. 13. The CI for this combination in SKI-DLCL cells is 0.646.

Example  5

We also examined the cytotoxic effects of the combination of IC20 (APL) and various doses of AraC on CRF-CEM and SKI-DLCL cell lines. Aplidine in both cell lines enhanced the effect of AraC and the IC50 of AraC. Dose was reduced from 30 nM to 1.6 nM in SKI-DLCL cell line, respectively, and from 10 nM to 0.8 nM in CCRF-CEM cell line (FIG. 14). Data was obtained using XTT assay after cell culture for 96 hours. The results represent the average of three different experiments.

Example 6 In vivo Study

We conducted an in vivo experiment to study the effects of aplidine alone and in combination with other drugs on lymphoid malignancies.

C. B.-17 scid Of scid  ( SCID  Permissible dose in mice) MTD Determination of

We used an in vivo model of human lymphoma in SCID mice for this purpose. Specifically, we used CCRF-CEMS cells and CB.17 scid / scid mice. We have experienced this model and have evaluated drug treatment using this xenograft (Lacerda J.F. et al. Blood 85 (10): 2675-2679 (1995)). We have found that the total dose of 1 mg / kg / week given in 5 daily doses is the maximum aplidine dose that can be tolerated by mice.

SCID  Single Agent and a Mouse Xenograft Model AraC Aplidine as a combination with Phosphorus  Determination of Vivo Antitumor Effects

10 ⁷ CEM-T leukemia cells were inoculated into SCID mice subcutaneously in the right flank. The mice were observed twice weekly for tumor formation at the site of inoculation. After palpable tumors were established, aplidine was injected in combination with a single agent and multiple doses of AraC to determine antitumor effects. Mice were randomized and administered aplidine and AraC combination for doses of 0.75 mg / kg and 1 mg / kg of aplidine alone, 50 mg / kg AraC alone, or all combinations of doses. The AraC dose selected for this combination is the dose at which the tumor growth was inhibited but no tumor regression occurred. All drugs were administered intraperitoneally, the tumor size was compared to the control mice group which received no treatment, and the combination group was compared to the single drug treated tumor size.

The most effective combination was found to be AraC-50 mg / kg + aplidine-0.75 mg / kg (FIG. 15).

These findings regarding aplidine can be extended to aplidine analogs, derivatives and related compounds. For example, the present invention relates to compounds such as the compounds of WO 0202596 and to anticancer drugs, preferably anti-leukemia drugs or anti-lymphoma drugs, in particular methotrexate, cytosine arabinoside, mitoxantrone, vinblastine, methylprednisolone or doxorubicin To provide a combination.

Claims (19)

Use of the aplidine or aplidine analog in the manufacture of a medicament for treating primary and / or metastatic cancer by a combination therapy using an aplidine or aplidine analog in combination with another drug. The use of the aplidine or aplidine analog of claim 1, wherein the cancer to be treated is leukemia or lymphoma. The use of claim 1 or 2, wherein the aplidine or aplidine analog and the other drug form part of the same drug. The aplidine or aplidine according to any one of claims 1 to 3, wherein the aplidine or aplidine analog and the other drug are provided as separate drugs for administration at the same time or at different times. Use of analogues. The aplidine or apliline according to any one of claims 1 to 4, wherein the other drug is selected from the group consisting of methotrexate, cytosine arabinoside, mitoxantrone, vinblastine, methylprednisolone and doxorubicin. Use of Dean Analogs. Use according to any one of claims 1 to 5, wherein the aplidine or aplidine analog is aplidine. Effective for the treatment of cancer, which is administered to a patient in need thereof prior to, during or after administering a therapeutically effective amount of aplidine or aplidine analog and the aplidine or aplidine analog A method of treating primary and / or metastatic cancer, comprising administering a therapeutically effective amount of another drug. 8. The method of claim 7, wherein the cancer to be treated is leukemia or lymphoma. The method of claim 7 or 8, wherein the other drug is selected from the group consisting of methotrexate, cytosine arabinoside, mitoxantrone, vinblastine, methylprednisolone and doxorubicin. A method of increasing the therapeutic efficacy of a drug effective for treating cancer, the method comprising administering an amount of aplidine or an aplidine analog to a patient in need thereof before, during or after administration of another drug How to. The method of claim 10, wherein the cancer to be treated is leukemia or lymphoma. The method of claim 10 or 11, wherein the drug is selected from the group consisting of methotrexate, cytosine arabinoside, mitoxantrone, vinblastine, methylprednisolone and doxorubicin. A pharmaceutical composition comprising an aplidine or an aplidine analog and other drugs effective for the treatment of cancer. The pharmaceutical composition of claim 13, wherein the other drug is effective for treating leukemia and / or lymphoma. The pharmaceutical composition according to claim 13 or 14, wherein the drug is selected from the group consisting of methotrexate, cytosine arabinoside, mitoxantrone, vinblastine, methylprednisolone and doxorubicin. Treatment of cancer, including instructions for the use of each agent in a combination of aplidine or an aplidine analog, dosage forms of other drugs effective for the treatment of cancer, and treatment or prevention of cancer Kit for use in prevention. The kit of claim 16, wherein the kit is for use in the treatment or prevention of leukemia and / or lymphoma. 18. The kit of claim 16 or 17, wherein said other drug is selected from the group consisting of methotrexate, cytosine arabinoside, mitoxantrone, vinblastine, methylprednisolone and doxorubicin. Use of aplidine or an aplidine analog in the manufacture of a medicament for the treatment of chronic lymphocytic leukaemia.

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